There is no effective standard therapy for patients with advanced metastatic melanoma. We have shown that eugenol, is an anti-melanoma agent in vitro and in vivo. The aim of this exploratory proposal is to establish proof of principle that (i) the safest and most effective dose of eugenol will depend on the route through which it is dispensed and (ii) E2F1 is a eugenol target in melanoma cells. Our working hypothesis is that there is a biphasic response to eugenol-induced DNA damage in melanoma cells. We have proposed that following eugenol treatment DNA damage signaling occurs through ATM/chk2 pathway that leads to stabilization of E2F1. Further there is temporal separation of E2F1 responses to eugenol as a function of DNA damage. As an immediate response E2F1 transactivates ogg1 to repair DNA base damage caused by eugenol. However as DNA damage persists E2F1 transcription activity is inhibited. As a result expression of PCNA, an E2F1 target gene does not occur and consequently cells are arrested in S phase. We will also determine the most efficacious method to deliver eugenol using the 1205Lu xenograft model. In this aim we will also determine the safest (non-toxic) eugenol dose. Results of this exploratory proposal will determine whether eugenol can be developed further as an anti-melanoma agent. Significance of this proposal is that eugenol targets a deregulated pathway in melanoma and because E2F1 regulates several genes involved in cellular processes such as apoptosis and metastasis eugenol can impact these pathways. There are no therapeutic agents in the immediate pipeline for advanced melanoma. Completion of the goals of this study will lead to subsequent proposals (RO1) aimed at obtaining a better understanding of the mechanism of action of eugenol and its clinical utility in preclinical models of melanoma. PUBLIC HEALTH RELEVANCE: Due to the lack of effective treatment options for metastatic malignant melanoma death rate continues to climb. The goal of the current proposal is to determine the most effective way to administer the compound and understand the mechanism of its growth inhibition activity. Completion of this project will take us closer to developing strategies to prevent fatal metastatic melanoma.